吉富罗非鱼肝脏蛋白质组双向电泳技术的建立与优化

Establishment and Optimization Analysis of Two-dimensional Electrophoresis System on GIFT Tilapia Liver

以吉富罗非鱼为研究对象,通过对其肝脏蛋白质的提取方法、等电聚焦程序和IPG胶条pH选择这几个关键环节的优化探索,建立了吉富罗非鱼肝脏蛋白质组双向电泳技术体系.结果表明：①采用液氮研磨法比匀浆器匀浆法提取蛋白质的效果更好;②聚焦时延长除盐时间至10 h,同时升高电压至8000 V,可获得清晰的电泳图谱;③通过双向电泳检测到吉富罗非鱼肝脏的蛋白点主要分布于pH值4.0 ～7.0之间.该技术体系的建立为今后开展罗非鱼蛋白质组学的进一步研究奠定了基础.

In the present study,genetic improvement of farmed tilapia （GIFT） was used as study material in the experiment.The two-dimensional electrophoresis （2-DE） for GIFT liver was established and optimized through comparative tests by using different extraction methods,isoelectric focusing programs and immobilized pH gradients （IPG） strips.The results showed that （i） the effect of extraction protein liquid nitrogen grinding was better than the homogenate machine; （ii） the desalting time added up to10 h and the voltage rose to 8000 V,which could get more clean two-dimensional electrophoresis map;（iii） The proteins of GIFT liver were obtained in pH 4.0-7.0 by 2-DE detection.So a two dimensional electrophoresis workflow for GIFT tilapia proteins separation had been establishment,which could laid a foundation for the further study on differential proteomics of tilapia.